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. 1996 Feb 2;132(4):667–679. doi: 10.1083/jcb.132.4.667

Selective stabilization of tau in axons and microtubule-associated protein 2C in cell bodies and dendrites contributes to polarized localization of cytoskeletal proteins in mature neurons

PMCID: PMC2199865  PMID: 8647897

Abstract

In mature neurons, tau is abundant in axons, whereas microtubule- associated protein 2 (MAP2) and MAP2C are specifically localized in dendrites. Known mechanisms involved in the compartmentalization of these cytoskeletal proteins include the differential localization of mRNA (MAP2 mRNA in dendrites, MAP2C mRNA in cell body, and Tau mRNA in proximal axon revealed by in situ hybridization) (Garner, C.C., R.P. Tucker, and A. Matus. 1988. Nature (Lond.). 336:674-677; Litman, P., J. Barg, L. Rindzooski, and I. Ginzburg. 1993. Neuron. 10:627-638), suppressed transit of MAP2 into axons (revealed by cDNA transfection into neurons) (Kanai, Y., and N. Hirokawa. 1995. Neuron. 14:421-432), and differential turnover of MAP2 in axons vs dendrites (Okabe, S., and N. Hirokawa. 1989. Proc. Natl. Acad. Sci. USA. 86:4127-4131). To investigate whether differential turnover of MAPs contributes to localization of other major MAPs in general, we microinjected biotinylated tau, MAP2C, or MAP2 into mature spinal cord neurons in culture (approximately 3 wk) and then analyzed their fates by antibiotin immunocytochemistry. Initially, each was detected in axons and dendrites, although tau persisted only in axons, whereas MAP2C and MAP2 were restricted to cell bodies and dendrites. Injected MAP2C and MAP2 bound to dendritic microtubules more firmly than to microtubules in axons, while injected tau bound to axonal microtubules more firmly than to microtubules in dendrites. Thus, beyond contributions from mRNA localization and selective axonal transport, compartmentalization of each of the three major MAPs occurs through local differential turnover.

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Selected References

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  1. Baumann K., Mandelkow E. M., Biernat J., Piwnica-Worms H., Mandelkow E. Abnormal Alzheimer-like phosphorylation of tau-protein by cyclin-dependent kinases cdk2 and cdk5. FEBS Lett. 1993 Dec 28;336(3):417–424. doi: 10.1016/0014-5793(93)80849-p. [DOI] [PubMed] [Google Scholar]
  2. Bernhardt R., Matus A. Light and electron microscopic studies of the distribution of microtubule-associated protein 2 in rat brain: a difference between dendritic and axonal cytoskeletons. J Comp Neurol. 1984 Jun 20;226(2):203–221. doi: 10.1002/cne.902260205. [DOI] [PubMed] [Google Scholar]
  3. Binder L. I., Frankfurter A., Rebhun L. I. The distribution of tau in the mammalian central nervous system. J Cell Biol. 1985 Oct;101(4):1371–1378. doi: 10.1083/jcb.101.4.1371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brion J. P., Guilleminot J., Couchie D., Flament-Durand J., Nunez J. Both adult and juvenile tau microtubule-associated proteins are axon specific in the developing and adult rat cerebellum. Neuroscience. 1988 Apr;25(1):139–146. doi: 10.1016/0306-4522(88)90013-9. [DOI] [PubMed] [Google Scholar]
  5. Brugg B., Matus A. Phosphorylation determines the binding of microtubule-associated protein 2 (MAP2) to microtubules in living cells. J Cell Biol. 1991 Aug;114(4):735–743. doi: 10.1083/jcb.114.4.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Caceres A., Binder L. I., Payne M. R., Bender P., Rebhun L., Steward O. Differential subcellular localization of tubulin and the microtubule-associated protein MAP2 in brain tissue as revealed by immunocytochemistry with monoclonal hybridoma antibodies. J Neurosci. 1984 Feb;4(2):394–410. doi: 10.1523/JNEUROSCI.04-02-00394.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen J., Kanai Y., Cowan N. J., Hirokawa N. Projection domains of MAP2 and tau determine spacings between microtubules in dendrites and axons. Nature. 1992 Dec 17;360(6405):674–677. doi: 10.1038/360674a0. [DOI] [PubMed] [Google Scholar]
  8. Cleveland D. W. Microtubule MAPping. Cell. 1990 Mar 9;60(5):701–702. doi: 10.1016/0092-8674(90)90083-q. [DOI] [PubMed] [Google Scholar]
  9. Cumming R., Burgoyne R. D., Lytton N. A. Immunofluorescence distribution of alpha tubulin, beta tubulin and microtubule-associated protein 2 during in vitro maturation of cerebellar granule cell neurones. Neuroscience. 1984 Jul;12(3):775–782. doi: 10.1016/0306-4522(84)90169-6. [DOI] [PubMed] [Google Scholar]
  10. Dotti C. G., Banker G. A. Experimentally induced alteration in the polarity of developing neurons. Nature. 1987 Nov 19;330(6145):254–256. doi: 10.1038/330254a0. [DOI] [PubMed] [Google Scholar]
  11. Drewes G., Lichtenberg-Kraag B., Döring F., Mandelkow E. M., Biernat J., Goris J., Dorée M., Mandelkow E. Mitogen activated protein (MAP) kinase transforms tau protein into an Alzheimer-like state. EMBO J. 1992 Jun;11(6):2131–2138. doi: 10.1002/j.1460-2075.1992.tb05272.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Garner C. C., Tucker R. P., Matus A. Selective localization of messenger RNA for cytoskeletal protein MAP2 in dendrites. Nature. 1988 Dec 15;336(6200):674–677. doi: 10.1038/336674a0. [DOI] [PubMed] [Google Scholar]
  13. Goedert M., Cohen E. S., Jakes R., Cohen P. p42 MAP kinase phosphorylation sites in microtubule-associated protein tau are dephosphorylated by protein phosphatase 2A1. Implications for Alzheimer's disease [corrected]. FEBS Lett. 1992 Nov 2;312(1):95–99. doi: 10.1016/0014-5793(92)81418-l. [DOI] [PubMed] [Google Scholar]
  14. Goedert M., Crowther R. A., Garner C. C. Molecular characterization of microtubule-associated proteins tau and MAP2. Trends Neurosci. 1991 May;14(5):193–199. doi: 10.1016/0166-2236(91)90105-4. [DOI] [PubMed] [Google Scholar]
  15. Gorbsky G. J., Sammak P. J., Borisy G. G. Chromosomes move poleward in anaphase along stationary microtubules that coordinately disassemble from their kinetochore ends. J Cell Biol. 1987 Jan;104(1):9–18. doi: 10.1083/jcb.104.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Graessmann A., Graessmann M., Mueller C. Microinjection of early SV40 DNA fragments and T antigen. Methods Enzymol. 1980;65(1):816–825. doi: 10.1016/s0076-6879(80)65076-9. [DOI] [PubMed] [Google Scholar]
  17. Grafstein B., Forman D. S. Intracellular transport in neurons. Physiol Rev. 1980 Oct;60(4):1167–1283. doi: 10.1152/physrev.1980.60.4.1167. [DOI] [PubMed] [Google Scholar]
  18. Harada A., Oguchi K., Okabe S., Kuno J., Terada S., Ohshima T., Sato-Yoshitake R., Takei Y., Noda T., Hirokawa N. Altered microtubule organization in small-calibre axons of mice lacking tau protein. Nature. 1994 Jun 9;369(6480):488–491. doi: 10.1038/369488a0. [DOI] [PubMed] [Google Scholar]
  19. Hirokawa N., Bloom G. S., Vallee R. B. Cytoskeletal architecture and immunocytochemical localization of microtubule-associated proteins in regions of axons associated with rapid axonal transport: the beta,beta'-iminodipropionitrile-intoxicated axon as a model system. J Cell Biol. 1985 Jul;101(1):227–239. doi: 10.1083/jcb.101.1.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hirokawa N., Hisanaga S., Shiomura Y. MAP2 is a component of crossbridges between microtubules and neurofilaments in the neuronal cytoskeleton: quick-freeze, deep-etch immunoelectron microscopy and reconstitution studies. J Neurosci. 1988 Aug;8(8):2769–2779. doi: 10.1523/JNEUROSCI.08-08-02769.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hirokawa N. Microtubule organization and dynamics dependent on microtubule-associated proteins. Curr Opin Cell Biol. 1994 Feb;6(1):74–81. doi: 10.1016/0955-0674(94)90119-8. [DOI] [PubMed] [Google Scholar]
  22. Hirokawa N., Shiomura Y., Okabe S. Tau proteins: the molecular structure and mode of binding on microtubules. J Cell Biol. 1988 Oct;107(4):1449–1459. doi: 10.1083/jcb.107.4.1449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Joshi H. C., Cleveland D. W. Differential utilization of beta-tubulin isotypes in differentiating neurites. J Cell Biol. 1989 Aug;109(2):663–673. doi: 10.1083/jcb.109.2.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kanai Y., Hirokawa N. Sorting mechanisms of tau and MAP2 in neurons: suppressed axonal transit of MAP2 and locally regulated microtubule binding. Neuron. 1995 Feb;14(2):421–432. doi: 10.1016/0896-6273(95)90298-8. [DOI] [PubMed] [Google Scholar]
  25. Kanai Y., Takemura R., Oshima T., Mori H., Ihara Y., Yanagisawa M., Masaki T., Hirokawa N. Expression of multiple tau isoforms and microtubule bundle formation in fibroblasts transfected with a single tau cDNA. J Cell Biol. 1989 Sep;109(3):1173–1184. doi: 10.1083/jcb.109.3.1173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Knops J., Kosik K. S., Lee G., Pardee J. D., Cohen-Gould L., McConlogue L. Overexpression of tau in a nonneuronal cell induces long cellular processes. J Cell Biol. 1991 Aug;114(4):725–733. doi: 10.1083/jcb.114.4.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kobayashi S., Ishiguro K., Omori A., Takamatsu M., Arioka M., Imahori K., Uchida T. A cdc2-related kinase PSSALRE/cdk5 is homologous with the 30 kDa subunit of tau protein kinase II, a proline-directed protein kinase associated with microtubule. FEBS Lett. 1993 Dec 6;335(2):171–175. doi: 10.1016/0014-5793(93)80723-8. [DOI] [PubMed] [Google Scholar]
  28. Kosik K. S., Finch E. A. MAP2 and tau segregate into dendritic and axonal domains after the elaboration of morphologically distinct neurites: an immunocytochemical study of cultured rat cerebrum. J Neurosci. 1987 Oct;7(10):3142–3153. doi: 10.1523/JNEUROSCI.07-10-03142.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kristofferson D., Mitchison T., Kirschner M. Direct observation of steady-state microtubule dynamics. J Cell Biol. 1986 Mar;102(3):1007–1019. doi: 10.1083/jcb.102.3.1007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ledesma M. D., Correas I., Avila J., Díaz-Nido J. Implication of brain cdc2 and MAP2 kinases in the phosphorylation of tau protein in Alzheimer's disease. FEBS Lett. 1992 Aug 17;308(2):218–224. doi: 10.1016/0014-5793(92)81278-t. [DOI] [PubMed] [Google Scholar]
  31. Lewis S. A., Ivanov I. E., Lee G. H., Cowan N. J. Organization of microtubules in dendrites and axons is determined by a short hydrophobic zipper in microtubule-associated proteins MAP2 and tau. Nature. 1989 Nov 30;342(6249):498–505. doi: 10.1038/342498a0. [DOI] [PubMed] [Google Scholar]
  32. Litman P., Barg J., Rindzoonski L., Ginzburg I. Subcellular localization of tau mRNA in differentiating neuronal cell culture: implications for neuronal polarity. Neuron. 1993 Apr;10(4):627–638. doi: 10.1016/0896-6273(93)90165-n. [DOI] [PubMed] [Google Scholar]
  33. Mandelkow E., Mandelkow E. M. Microtubules and microtubule-associated proteins. Curr Opin Cell Biol. 1995 Feb;7(1):72–81. doi: 10.1016/0955-0674(95)80047-6. [DOI] [PubMed] [Google Scholar]
  34. Mandell J. W., Banker G. A. The microtubule cytoskeleton and the development of neuronal polarity. Neurobiol Aging. 1995 May-Jun;16(3):229–238. doi: 10.1016/0197-4580(94)00164-v. [DOI] [PubMed] [Google Scholar]
  35. Matsuo E. S., Shin R. W., Billingsley M. L., Van deVoorde A., O'Connor M., Trojanowski J. Q., Lee V. M. Biopsy-derived adult human brain tau is phosphorylated at many of the same sites as Alzheimer's disease paired helical filament tau. Neuron. 1994 Oct;13(4):989–1002. doi: 10.1016/0896-6273(94)90264-x. [DOI] [PubMed] [Google Scholar]
  36. Meichsner M., Doll T., Reddy D., Weisshaar B., Matus A. The low molecular weight form of microtubule-associated protein 2 is transported into both axons and dendrites. Neuroscience. 1993 Jun;54(4):873–880. doi: 10.1016/0306-4522(93)90581-y. [DOI] [PubMed] [Google Scholar]
  37. Nixon R. A., Fischer I., Lewis S. E. Synthesis, axonal transport, and turnover of the high molecular weight microtubule-associated protein MAP 1A in mouse retinal ganglion cells: tubulin and MAP 1A display distinct transport kinetics. J Cell Biol. 1990 Feb;110(2):437–448. doi: 10.1083/jcb.110.2.437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Okabe S., Hirokawa N. Microtubule dynamics in nerve cells: analysis using microinjection of biotinylated tubulin into PC12 cells. J Cell Biol. 1988 Aug;107(2):651–664. doi: 10.1083/jcb.107.2.651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Okabe S., Hirokawa N. Rapid turnover of microtubule-associated protein MAP2 in the axon revealed by microinjection of biotinylated MAP2 into cultured neurons. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4127–4131. doi: 10.1073/pnas.86.11.4127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Papasozomenos S. C., Binder L. I., Bender P. K., Payne M. R. Microtubule-associated protein 2 within axons of spinal motor neurons: associations with microtubules and neurofilaments in normal and beta,beta'-iminodipropionitrile-treated axons. J Cell Biol. 1985 Jan;100(1):74–85. doi: 10.1083/jcb.100.1.74. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Papasozomenos S. C., Binder L. I. Phosphorylation determines two distinct species of Tau in the central nervous system. Cell Motil Cytoskeleton. 1987;8(3):210–226. doi: 10.1002/cm.970080303. [DOI] [PubMed] [Google Scholar]
  42. Paudel H. K., Lew J., Ali Z., Wang J. H. Brain proline-directed protein kinase phosphorylates tau on sites that are abnormally phosphorylated in tau associated with Alzheimer's paired helical filaments. J Biol Chem. 1993 Nov 5;268(31):23512–23518. [PubMed] [Google Scholar]
  43. Peng I., Binder L. I., Black M. M. Biochemical and immunological analyses of cytoskeletal domains of neurons. J Cell Biol. 1986 Jan;102(1):252–262. doi: 10.1083/jcb.102.1.252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Ransom B. R., Neale E., Henkart M., Bullock P. N., Nelson P. G. Mouse spinal cord in cell culture. I. Morphology and intrinsic neuronal electrophysiologic properties. J Neurophysiol. 1977 Sep;40(5):1132–1150. doi: 10.1152/jn.1977.40.5.1132. [DOI] [PubMed] [Google Scholar]
  45. Sabry J., O'Connor T. P., Kirschner M. W. Axonal transport of tubulin in Ti1 pioneer neurons in situ. Neuron. 1995 Jun;14(6):1247–1256. doi: 10.1016/0896-6273(95)90271-6. [DOI] [PubMed] [Google Scholar]
  46. Sato-Yoshitake R., Shiomura Y., Miyasaka H., Hirokawa N. Microtubule-associated protein 1B: molecular structure, localization, and phosphorylation-dependent expression in developing neurons. Neuron. 1989 Aug;3(2):229–238. doi: 10.1016/0896-6273(89)90036-6. [DOI] [PubMed] [Google Scholar]
  47. Shiomura Y., Hirokawa N. Colocalization of microtubule-associated protein 1A and microtubule-associated protein 2 on neuronal microtubules in situ revealed with double-label immunoelectron microscopy. J Cell Biol. 1987 Jun;104(6):1575–1578. doi: 10.1083/jcb.104.6.1575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Shiomura Y., Hirokawa N. The molecular structure of microtubule-associated protein 1A (MAP1A) in vivo and in vitro. An immunoelectron microscopy and quick-freeze, deep-etch study. J Neurosci. 1987 May;7(5):1461–1469. doi: 10.1523/JNEUROSCI.07-05-01461.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Takeuchi M., Hisanaga S., Umeyama T., Hirokawa N. The 72-kDa microtubule-associated protein from porcine brain. J Neurochem. 1992 Apr;58(4):1510–1516. doi: 10.1111/j.1471-4159.1992.tb11372.x. [DOI] [PubMed] [Google Scholar]
  50. Tucker R. P., Binder L. I., Matus A. I. Neuronal microtubule-associated proteins in the embryonic avian spinal cord. J Comp Neurol. 1988 May 1;271(1):44–55. doi: 10.1002/cne.902710106. [DOI] [PubMed] [Google Scholar]
  51. Tytell M., Brady S. T., Lasek R. J. Axonal transport of a subclass of tau proteins: evidence for the regional differentiation of microtubules in neurons. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1570–1574. doi: 10.1073/pnas.81.5.1570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Vallee R. B. A taxol-dependent procedure for the isolation of microtubules and microtubule-associated proteins (MAPs). J Cell Biol. 1982 Feb;92(2):435–442. doi: 10.1083/jcb.92.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Vulliet R., Halloran S. M., Braun R. K., Smith A. J., Lee G. Proline-directed phosphorylation of human Tau protein. J Biol Chem. 1992 Nov 5;267(31):22570–22574. [PubMed] [Google Scholar]
  54. Wiche G., Oberkanins C., Himmler A. Molecular structure and function of microtubule-associated proteins. Int Rev Cytol. 1991;124:217–273. doi: 10.1016/s0074-7696(08)61528-4. [DOI] [PubMed] [Google Scholar]

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